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Related Concept Videos

Affinity Chromatography01:03

Affinity Chromatography

Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
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Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
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Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization
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Affinity-based chemoproteomics with small molecule-peptide conjugates.

Chaitanya Saxena1

  • 1Shantani Proteome Analytics Pvt. Ltd., Pune, MH, India. csaxena@shantani.com

Methods in Molecular Biology (Clifton, N.J.)
|November 9, 2011
PubMed
Summary

Small molecule-peptide conjugates (SMPCs) overcome limitations in chemoproteomics by preserving probe activity for target protein identification. This method enhances the capture and analysis of small molecule-protein interactions in complex biological samples.

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Last Updated: May 27, 2026

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Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
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Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

Published on: July 31, 2011

Area of Science:

  • Chemical Biology
  • Proteomics
  • Molecular Biology

Background:

  • Direct immobilization of small bioactive probes in chemoproteomics can reduce their functional activity.
  • Immobilized probes often show lower affinity for target proteins, hindering specific capture and leading to losses.
  • Existing methods face challenges in preserving probe efficacy and achieving specific protein target identification.

Purpose of the Study:

  • To develop a novel strategy for capturing protein targets of small molecules.
  • To overcome the limitations of direct probe immobilization in affinity-based chemoproteomics.
  • To enable the identification of small molecule targets from complex biological samples like cell lysates and intact cells.

Main Methods:

  • Design and synthesis of small molecule-peptide conjugates (SMPCs).
  • Utilizing peptides with tailored biochemical and biophysical properties.
  • Application of SMPCs for capturing protein targets from cell lysates and intact cells.
  • Labeling SMPCs with fluorophores for cellular localization studies.

Main Results:

  • SMPCs effectively preserve the functional activity of small molecule probes.
  • Successful identification of protein targets using SMPCs from various biological sources.
  • Demonstrated ability to capture targets from both cell lysates and intact cells.
  • Fluorophore-labeled SMPCs provided insights into target protein cellular distribution.

Conclusions:

  • Small molecule-peptide conjugates represent a versatile platform for chemoproteomics.
  • SMPCs overcome the activity loss associated with direct probe immobilization.
  • This approach facilitates robust identification and localization of small molecule-protein interactions.